Alkaloids : An Organic Chemistry

Perspective
AMALIN HUSNA BINTI MOHD AZHAR
YAP MING YAN
MARCELLA MARY A/P RICHARD
BHUVANESWARI A/P RAGUNATHAN

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 Alkaloids are traditionally basic nitrogen-
containing organic constituents that occur
mainly in plants. Alkaloids often have
pronounced bioactivities and are therefore
thought to play an important role in the
interaction of plants with their environment.
The purpose of the project is to gain a deeper
knowledge about the significance of alkaloids
occurring in the plants surrounding us .
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 Which part(s) of the plant samples have the highest
amount of alkaloids?
Which way(s) of sample preparation will give more
amount of alkaloids to be extracted in plants?
How to extract alkaloids from plant samples?
What is the change of the mixture after the Mayers
reagent is added in the extract of the plant if alkaloid is
present?
How does one species of plants have more alkaloid
content than the other?

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 To identify the presence of alkaloids in plant
parts
To measure the percentage of composition of
alkaloids in different plant parts

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 The project is carried out in two parts :
Part A : Phytochemical screening of alkaloids
-Method X: Boiling distilled water + HCL
-Method Y: Ammoniacal chloroform
Part B : Acid-base extraction of alkaloids

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 Part A : Phytochemical screening
Method X : Boiling distilled water with crushed
sample + HCl
150 mL Beaker - Glass rod
Wire gauze - 3 Test tubes
Bunsen burner - Dropper
Tripod stand - 50 mL distilled water
Electronic weight balance - 2g of plant sample
150 mL Conical flask - Mayers reagent
Filter funnel - 5mL 2.0 M Hydrochloric
Filter paper acid
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 Part A : Phytochemical screening
Method Y : Ammoniacal chloroform with crushed
sample
150 mL Beaker - Dropper
Electronic weight balance - 2g of plant sample
150 mL Conical flask - 50 mL Ammoniacal
Filter funnel chloroform
- Mayers reagent
Filter paper
- 2.0 M Sulphuric acid
Glass rod
6 Test tubes
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 2 grams of crushed sample are weighed. 50 mL of distilled water is
added inside the beaker with the crushed sample.
Bunsen burner is used to heat the beaker until boiling, for at least 2
minutes. Then, the heat is turned off and the beaker is left for
cooling for another 2 minutes.
Filter funnel with loosely packed cotton and filter paper is used to
filter the mixture.
5 mL of 2.0 M of hydrochloric acid, HCl is added to the filtrate and
the mixture is swirl to ensure the solution is well mixed.
The filtrate is separated into 3 test tubes. One test tube is being
kept as a reference.
2 to 3 drops of Mayers reagent is added to one of the test tube.
Observation is recorded. Alkaloid is present if cream precipitate
formed or the mixture turns cloudy.

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 30 ml of ammoniacal chloroform are added to 2 g of the crushed sample
(Ammoniacal chloroform is 1% NH3 chloroform; prepared by shaking 1ml
concentrated ammonia in 1L chloroform).
The mixture is stir to form a loose paste. Then, the mixture is filtered with
a bit of loosely packed cotton and filter paper using the filter funnel.
20 mL of ammoniacal chloroform is added in the filter funnel to rinse and
get more of the organic compunds in plant samples
20 to 40 drops of 2.0M of H2SO4 are added to the filtrate and gently shaken
to avoid emulsion forming.
Filtrate is divided into three test tubes. One test tube would be the
reference and the other two can be used further.
For the two layers to separate, wait for a day.
By using a dropper extract the organic layer (top layer) and transfer to a
clean test tube, 2 to 3 drops of Mayers reagent is added. alkaloid is present
if cream precipitate formed or the mixture turns into cloudy.
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 Part B : Acid - base extraction of alkaloids
2 Beakers (150 mL) - Glass rod
3 Beakers (120 mL) - 2 Test tubes
Electronic weight balance - Dropper
- 2g of plant sample
Separating funnel
- 50 mL Ammoniacal
150 mL Conical flask chloroform
Filter funnel - Mayers reagent
Filter paper - 20 mL 1.0 M Sulphuric
acid
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 By using 50 mL of ammoniacal chloroform extract the dry sample just
like in phytochemical screening test and add 20ml of 1.0M sulphuric
acid to the filtrate.
The mixture is transferred to a separating funnel and shaken.
Extract and keep the aqueous layer as well as add 20 ml of 1.0M
sulphuric acid again after that on top of the organic layer and repeat.
Aqueous layer for presence of alkaloid is tested using Mayers reagent,
aqueous layer that are tested positive are kept.
Extraction is repeated using dilute acid until alkaloids are not detected
with Mayers reagent.
All the aqueous layers are put together and a base is added to the
aqueous layers such as solid sodium carbonate to neutralize the acid.
The mixture is tested with litmus paper to ensure it is neutral or
slightly alkaline.
The solution is heated in crucible to evaporate the water. Dry it further
at the fume chamber or the oven and leave for one day. The final
product is the refined extract of alkaloids.
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 Part A : Phytochemical screening
( Method X : Boiling + HCL )
Observation ( test with
Plant Sample Deduction
Mayers solution )
Colourless solution
Tobacco leaves Alkaloids present
turns cloudy
Colourless solution
Periwinkle leaves Alkaloids present
turns cloudy
Colourless solution
Hibiscus leaves Alkaloid absent
remains unchanged
Colourless solution
Papaya leaves Alkaloid present
turns cloudy
Colourless solution
Curry leaves Alkaloid present
turns cloudy
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 Part A : Phytochemical Screening of alkaloids
( Method Y : ammonical chloroform +
sulphuric acid )
Observations ( test with
Plant Samples Deduction
Mayers solution )
Colourless solution turns
Tobacco leaves Alkaloid present
cloudy
Colourless solution turns
Periwinkle leaves Alkaloid present
cloudy
Colourless solution remains
Hibiscus leaves Alkaloid absent
unchanged
Colourless solution turns
Papaya leaves Alkaloid present
cloudy
Colourless solution remains
Curry leaves Alkaloid absent
unchanged

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 Phytochemical screening for alkaloids
Tobacco, periwinkle and papaya leaf samples was tested positive
result with Mayers reagent on both of the phytochemical
screening methods.
However, curry leaves shows positive results in phytochemical
screening using boiling method when colourless test solution
reacts with Mayers solution forms cloudy solution but show
negative results in phytochemical screening using 1% ammonical
chloroform solution when colourless test solution reacts as
Mayers solution remain unchanged.
Hibiscus was tested negative in both methods. Inyang et al. in
Journal of Medicinal Plants Studies, said that alkaloids is present in
hibiscus leaf but in a low concentration. We suggested that hibiscus
leaf sample might actually have alkaloids but it is too low in
concentration that it could not be detected with Mayers reagent.

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1) All the base 2 3 are completely neutralise with the acid
2 4 , with 2 are given off
2 + (aq) + 32 (s) 2 (g) + 2 O ( l )

2) A small amount of the sulphuric acid solution ( extracted

colourless layer which contains alkaloid ) are transferred into a
test tube to react with Mayers solution to test the presence of
alkaloids . Eventually , part of the amount of alkaloid are lost .

3) During heating the neutralised mixture , the boiling of the

mixture are reactive and volatile , causing some of the liquid (
unable to vaporise smoothly ) escape from the crucible ,
resulting in lost of small amount of alkaloids that should be
obtained .
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 Phytochemical Screening of alkaloids (Method X)
No modifications made
Phytochemical Screening of alkaloid (Method Y)
A volume of 50 mL of ammoniacal chloroform was
added to 2 g of plant sample instead of the proposed
3 to 5 mL to get more organic compound of plant
sample
Additional 20 drops of 2.0 M sulphuric acid was added
(Total volume = 40 drops) to extract more alkaloids in
aqueous layer (top layer)
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 Acid Base Extraction of alkaloids
Use solid sodium carbonate instead of aqueous sodium
carbonate in order to eliminate excess water so that all base are
completely react with the acid use .
Bigger crucible are suggested to be used in order to reduce the
amount of loss of alkaloids escape through heating .
Allow the sample to be air dry instead of heating to reduce the
loss of alkaloids through boiling . If heating is used , low heating
is suggested as the solution heated is volatile and , thus some
alkaloid pops out from the crucible .

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 The project of investigation on alkaloids shows
that the sample plant leaves that show
presence of alkaloid ( positive results ) are
tobacco , periwinkle and papaya leaves
wheareas hibiscus leaves show negative result
for both phytochemical screening test
For the case of curry leaf , show positive result
for Method X but negative result for Method
Y.

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 Acid-base extraction is carried out using
tobacco leaves . The mass of the alkaloid
obtained ( 2.70 g ) is less than the theoretical
mass of the alkaloid calculated (2.84 g ).This is
due to the fact that part of the alkaloid salt
are lost during the process from extracting it .
The white solid obtained which consists of
salt , alkaloid salt , organic content and
impurities . Thus , the mass of the alkaloid
existed only less than but approximate to the
calculated theoretical mass .

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THANK YOU

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